Recombinant Human C17orf49 Protein: Handling & Usage Protocol

Recombinant Human C17orf49 Protein is a research-grade recombinant protein requiring careful reconstitution, controlled storage, and validated handling procedures to maintain full biological activity in downstream experimental applications.

Product Name

Recombinant Human C17orf49 Protein

SKU

rp365735

Reference Catalog

absea#PP-1353

Species Origin

Human (Homo sapiens)

Recommended Storage

Store at -20°C or -80°C; avoid repeated freeze-thaw cycles

Application

Research use only; suitable for in vitro assays including SDS-PAGE, Western blot, and functional studies

Materials and Equipment Required

Before beginning the protocol, ensure the following materials and equipment are available:

• Recombinant Human C17orf49 Protein (SKU: rp365735)
• Sterile deionized or ultrapure water (or buffer as specified in the product datasheet)
• Microcentrifuge tubes (low-protein-binding, 1.5 mL or 0.5 mL)
• Calibrated micropipettes and sterile filter tips
• Benchtop microcentrifuge
• Ice bucket with wet ice or dry ice for sample handling
• Biosafety cabinet (recommended for aseptic technique)
• -20°C and/or -80°C freezer for storage
• Vortex mixer (for gentle mixing only)

Note: Always consult the product-specific datasheet for buffer composition, protein concentration, and any special reconstitution requirements before proceeding.

Reconstitution Procedure

Follow these steps to reconstitute the lyophilized Recombinant Human C17orf49 Protein:

1. Equilibrate the vial: Remove the protein vial from -20°C or -80°C storage and allow it to equilibrate to room temperature (18–25°C) for 10–15 minutes before opening. This reduces condensation and minimizes protein aggregation.
2. Brief centrifugation: Briefly centrifuge the sealed vial at low speed (e.g., 500–1000 × g for 30 seconds) to collect lyophilized material at the bottom of the vial.
3. Add reconstitution buffer: Add the recommended volume of sterile reconstitution buffer or ultrapure water slowly along the side of the vial wall. Do not pipette directly onto the lyophilized cake to avoid foaming or denaturation.
4. Gentle mixing: Gently swirl or tap the vial to dissolve the protein. Avoid vigorous vortexing. If necessary, allow the vial to stand at room temperature for 5–10 minutes with occasional gentle mixing.
5. Verify dissolution: Visually confirm the solution is clear and free of particulates. If turbidity persists, centrifuge at low speed and use the supernatant.
6. Aliquot immediately: Prepare single-use aliquots in low-protein-binding tubes to minimize freeze-thaw cycles and degradation.

Important: The recommended reconstitution volume and buffer should be taken from the product datasheet (absea#PP-1353). Contact Aladdin Scientific technical support if the datasheet is unavailable.

Storage and Stability Guidelines

Proper storage of Recombinant Human C17orf49 Protein is critical for maintaining its integrity and activity:

• Lyophilized form: Store at -20°C in a desiccated environment, protected from light and moisture. Under optimal conditions, lyophilized protein is stable for 12 months or longer from the date of manufacture.
• Reconstituted protein: After reconstitution, if the protein will be used within 1–2 weeks, store aliquots at 4°C. For longer-term storage, freeze aliquots at -20°C or -80°C.
• Freeze-thaw cycles: Limit freeze-thaw cycles to an absolute minimum (no more than 2–3 cycles recommended). Each cycle risks partial denaturation, aggregation, and loss of biological activity.
• Carrier protein: Addition of a carrier protein such as 0.1% bovine serum albumin (BSA) to the reconstitution buffer may enhance stability during long-term storage, particularly at low protein concentrations. Verify compatibility with downstream assays before use.
• Do not store reconstituted protein at room temperature for extended periods.

Disclaimer: Specific stability data for C17orf49 protein should be confirmed from the product datasheet (absea#PP-1353). The guidelines above reflect general best practices for recombinant proteins.

Recommended Experimental Applications and Working Concentrations

Recombinant Human C17orf49 Protein (rp365735) is intended for research use only and is suitable for a range of in vitro applications. Typical applications include:

• SDS-PAGE and Western Blot: Use as a positive control or molecular weight standard for C17orf49 detection. Load 0.1–1 µg per lane as a starting point and optimize based on antibody sensitivity.
• ELISA development: Can be used as a coating antigen or standard in enzyme-linked immunosorbent assays targeting C17orf49.
• Antibody validation: Useful for confirming specificity and sensitivity of anti-C17orf49 antibodies in immunoassay development.
• Functional/biochemical assays: May be used to investigate protein–protein interactions or enzymatic activity, subject to the biological function attributed to C17orf49 in the literature.

Note on working concentrations: Optimal concentrations are assay-dependent and must be determined empirically by the end user. Always perform a titration experiment to establish the minimal effective concentration for your specific assay system.

Important: This product is not approved for use in clinical diagnostics, therapeutic applications, or administration to humans or animals.

Quality Control and Troubleshooting

To verify protein quality and troubleshoot common issues, follow these recommendations:

Quality Control Checks

• SDS-PAGE purity assessment: Run reconstituted protein on a denaturing SDS-PAGE gel to verify molecular weight and purity. The expected band should be consistent with the theoretical molecular weight of C17orf49.
• Protein quantification: Quantify protein concentration using a validated method such as BCA assay or Bradford assay after reconstitution to confirm yield before use.
• Visual inspection: Check for clarity of solution; turbid or particulate-containing solutions may indicate aggregation and should be discarded or centrifuged prior to use.

Common Issues and Solutions

• Protein aggregation: Caused by improper reconstitution, excessive freeze-thaw cycles, or inappropriate storage temperature. Solution: Reconstitute according to the datasheet, aliquot into single-use portions, and store at recommended temperatures.
• Low or no signal in assay: May be due to protein degradation, suboptimal antibody concentration, or incompatible assay conditions. Solution: Verify protein integrity by SDS-PAGE, re-optimize antibody concentrations, and check buffer compatibility.
• Unexpected band pattern: Could reflect proteolytic degradation or incorrect molecular weight expectations. Solution: Compare to the product datasheet and positive controls; ensure protease inhibitors are used where appropriate.

For additional technical assistance, contact Aladdin Scientific customer support and reference SKU rp365735 and catalog number absea#PP-1353.

Frequently asked questions

What is the recommended reconstitution buffer for Recombinant Human C17orf49 Protein?

The reconstitution buffer and volume are specified in the product datasheet (absea#PP-1353). Generally, sterile ultrapure water or a physiological buffer such as PBS (pH 7.4) is used. Always refer to the specific datasheet for the exact recommendation, as buffer requirements may vary by formulation.

How many freeze-thaw cycles can Recombinant Human C17orf49 Protein tolerate?

It is recommended to limit freeze-thaw cycles to no more than 2–3 times to preserve protein activity and integrity. To minimize exposure to freeze-thaw stress, aliquot the protein into single-use volumes immediately after reconstitution and store unused aliquots at -80°C.

Is Recombinant Human C17orf49 Protein suitable for in vivo or clinical use?

No. Recombinant Human C17orf49 Protein (rp365735) is strictly for research use only and is not approved, validated, or intended for use in clinical diagnostics, therapeutic procedures, or administration to humans or animals.

How should I verify the quality of the reconstituted protein before use?

Run the reconstituted protein on an SDS-PAGE gel to confirm the expected molecular weight band and assess purity. Additionally, quantify protein concentration using a BCA or Bradford assay, and visually inspect the solution for clarity. Any turbidity or unexpected banding pattern should be investigated before proceeding with downstream experiments.

Can carrier protein be added to improve the stability of the reconstituted C17orf49 protein?

Yes, adding a carrier protein such as 0.1% BSA (bovine serum albumin) to the reconstitution buffer is a common strategy to enhance protein stability during storage, particularly when working at low protein concentrations. However, verify that the carrier protein is compatible with your intended downstream assay before adding it.

References

1. Aladdin Scientific Product Datasheet: Recombinant Human C17orf49 Protein, absea#PP-1353, SKU rp365735.
2. General best practices for handling and storage of recombinant proteins — textbook-level laboratory guidance. Specific references to C17orf49 function and stability require confirmation from primary literature.

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.